Source tracing method for bacteria in infant formula milk powder

An infant formula and bacteria technology, applied in the field of food safety, can solve the problems of single detection and single prevention and control of bacteria, traceability of finished product bacteria source, pollution and other problems

Inactive Publication Date: 2016-10-12
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is precisely because of the non-absolute sterility in the processing process that infant formula milk powder may be contaminated by pathogenic bacteria from raw materials and production processes, which will have adverse effects on the health of infants and young children.
At present, the prevention and control of bacteria in infant formula milk powder is limited to a single detection and single prevention and control of finished products or raw materials and production environments, which cannot achieve the purpose of tracing the source of bacteria in finished products from the root and targeted prevention and control

Method used

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  • Source tracing method for bacteria in infant formula milk powder
  • Source tracing method for bacteria in infant formula milk powder
  • Source tracing method for bacteria in infant formula milk powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 sample collection

[0030] All the samples used in the research (raw material samples, air samples, surface coating samples) were collected from an infant formula milk powder processing plant in winter (November 2014) and summer (June 2015) by our laboratory staff respectively. factory. See Table 3 for the sample information of raw and auxiliary materials, and three samples were collected for each type of sample.

[0031] Table 3 Raw and auxiliary materials of infant formula milk powder

[0032]

[0033] For air sample and surface smear sample collection areas, see infant formula milk powder processing flow chart figure 1 , a total of 6 sampling areas, including 5 indoor areas and 1 outdoor area. Among them, the indoor area covers the main links of the infant formula milk powder processing process, including the pre-treatment link (A), the production link (B), the production in-package link (C), the packaging in-package link (D), and the packaging link (...

Embodiment 2

[0036] The separation of embodiment 2 bacteria:

[0037] Samples of raw materials

[0038] Weigh 25g of raw and auxiliary material samples of infant formula milk powder into a sterilized 225mL BPW Erlenmeyer flask, mix thoroughly, take 1mL of the mixed solution and add it to 9mL sterile PBS, after mixing, take 1mL and add it to another 9mL sterile In PBS, carry out gradient dilution by analogy, select the appropriate dilution gradient, absorb 100 μL of the mixed solution for each dilution gradient, and evenly spread it on the TSA solid medium, each sample is 3 parallel, and cultivate it in a constant temperature incubator at 37°C for 24 hours , select a single colony with different appearances in LB liquid medium, mix them and culture them in a 37°C incubator for 24h. Purification is required for non-single colonies. First, use the inoculation loop to dip the shaken overnight cultured LB bacterial solution, line the TSA solid medium with three zones, and incubate it upside do...

Embodiment 3

[0042] Embodiment 3 bacterial identification:

[0043] Genomic DNA Extraction

[0044] For the raw material samples of infant formula milk powder and surface smear samples, 2 mL of LB liquid bacterial liquid of the isolated strains were taken respectively, and the DNA of the isolated strains was extracted according to the specific method required by the TIANamp Bacterial Genomic DNA Extraction Kit.

[0045] Utilize bacterial genomic DNA extraction kit to carry out DNA extraction to isolated bacteria, and detect on 1% agarose gel electrophoresis, some results are as follows figure 2 shown.

[0046] It can be seen from the figure that the extracted DNA of the isolated strain has only one clear band without interference from other bands, indicating that the DNA extraction results meet the requirements and can be used for subsequent amplification experiments.

[0047] PCR amplification system and cycle conditions

[0048](1) PCR amplification of 16S rDNA sequence

[0049] Amp...

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Abstract

The invention provides a source tracing method for bacteria in infant formula milk powder. The source tracing method comprises the following steps: separating bacteria in raw materials and auxiliary materials of the infant formula milk powder and in the production environment; carrying out DNA extraction on the separated strains; carrying out PCR amplification on the 16S rDNA sequence by utilizing universal primers FA-27F: 5'-AGTCTCTGATCATGCCTCAG-3' and RA-1492R: 5'-AAGGAGGTGCTCCAGCC-3', and detecting the amplification product on 1% of agarose gel electrophoresis; sequencing the PCR amplification product of the 16S rDNA sequence obtained in the step (3); establishing a phylogenetic tree by adopting the obtained 16S rDNA gene sequence of the strains and by utilizing MEGA software in DNAStar, and analyzing the phylogenetic tree to achieve the purpose of tracing the source of the bacteria in the infant formula milk powder.

Description

technical field [0001] The invention belongs to the field of food safety, and in particular relates to a method for traceability of bacteria in formula milk powder for infants and young children. Background technique [0002] As the primary substitute of breast milk, infant formula milk powder is an important source of food intake for infants, providing almost all protein, lipids, vitamins, minerals and other nutrients for the healthy growth and development of infants. Infant formula milk powder is required to be a product free of harmful bacteria, but the raw and auxiliary materials used in its production are prone to contamination of bacteria, and its production process is not an absolutely sterile environment. It is precisely because of the non-absolute sterility in the processing process that infant formula milk powder may be contaminated by pathogenic bacteria from raw materials and production processes, which will have adverse effects on the health of infants and young...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/689C12Q1/686
Inventor 姜毓君满朝新娄彬彬潘瑞丽周彦宏
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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